摘要
利用甲基磺酸乙酯(EMS)化学诱变单季常规晚粳稻浙粳99,获得叶早衰突变体es33。为揭示es33突变体早衰的分子调控机制,以野生型浙粳99为对照,对突变体进行表型鉴定,统计农艺性状和测定光合性能,并对早衰基因ES33进行定位。结果表明,突变体es33幼苗在播种后第10天,第2和第3叶的叶尖出现明显干枯早衰现象,与野生型相比,分蘖盛期叶片光合色素含量降低、叶绿体结构疏松、光合作用减弱、部分酶活性降低,成熟期植株矮小、分蘖少、衰老严重。遗传分析结果表明,突变体es33早衰性状受一对隐性核基因控制,利用基因定位和重测序技术,将控制该性状的基因定位在水稻第3号染色体上的SSR标记STS-15和G24之间,物理距离为977.8 kb,经测序确定LOC_Os03g31550为候选基因;ES33基因第11个外显子发生4个碱基的缺失,造成移码突变,导致蛋白(黄嘌呤脱氢酶)翻译提前终止;通过系统发育树分析和氨基酸序列比对得知,ES33蛋白在禾本科作物中高度保守。本研究结果为进一步探究早衰基因ES33的功能奠定了基础。
The es33(early senescence 33)mutant was isolated from a photosensitive japonica rice cultivar Zhejing 99 with Ethyl Methane Sulfonate(EMS)treatment.In order to reveal the molecular regulation mechanism of ES33,we characterized and cloned the gene responsible for the es33 mutation.Results showed that the es33 mutant exhibited a leaf tip dryness and premature senescence phenotype at the second and third leaf tip after sowing 10 days showed significantly reduced levels of photosynthetic pigment,impaired chloroplast structure,lowered capacity of photosynthesis,reduced enzyme activities at tillering stage,and dwarf plants,few tillers,and severe senescence at maturing stage compared with the wild-type.Genetic analysis indicated that the early senescence phenotype was controlled by a single recessive nuclear gene,which was located between SSR markers STS-15 and G24 on chromosome 3 with a physical distance of 977.8 kb by using gene mapping and resequencing technology.Sequencing confirmed that LOC_Os03g31550 was a candidate gene,a bases deletion in its 11 th exon was detected in mutant,which resulting in a frameshift mutation lead to premature termination of protein(xanthine dehydrogenase)translation.Phylogenetic tree analysis and amino acid sequence alignment showed that ES33 was highly conserved in gramineous crops.The results laid a foundation for further study of the function of early senescence gene ES33.
作者
郭铧艳
叶胜海
翟荣荣
朱国富
俞法明
巫明明
叶靖
张小明
GUO Huayan;YE Shenghai;ZHAI Rongrong;ZHU Guofu;YU Faming;WU Mingming;YE Jing;ZHANG Xiaoming(College of Chemistry and Life Science,Zhejiang Normal University,Jinhua,Zhejiang 321004;Institute of Crop and Nuclear Technology Utilization,Zhejiang Academy of Agricultural Science,Hangzhou,Zhejiang 310021)
出处
《核农学报》
CAS
CSCD
北大核心
2022年第12期2317-2329,共13页
Journal of Nuclear Agricultural Sciences
基金
浙江省公益技术研究计划项目(LGN20C130005)
浙江省重点研发计划项目(2021C02063-5)。
